BioLux Cypridina Luciferase Assay Buffer (1X) and Substrate (lyophilized powder) must always be protected from light.
BioLux Cypridina Luciferase Substrate is supplied in lyophilized form to ensure long shelf life and maximum stability.
Usage notes:
Store the BioLux Cypridina Luciferase Assay Buffer (1X) at -20°C for up to 1 year.
The BioLux Cypridina Luciferase Assay Buffer (1X), the reconstituted CLuc substrate (100X solution) and the CLuc assay solution must be protected from light.
The CLuc assay solution should be freshly prepared and incubated at room temperature for 30 minutes (protect from light) before use.
The linear range of the luminometer must be established. This can be easily done by assaying serial dilutions of a CLuc containing sample (Figure 3), such as the culture medium of CLuc-expressing cells. In addition, the assay solution itself as well as the conditioned medium (culture medium from untransfected cells) should be included to establish the background in the assay.
If excess activity for the instrument range is found, the sample should be diluted in either PBS or 10% serum-containing medium. The integration time can also be reduced, e.g. 2 seconds instead of 5 seconds.
The presence of serum, i.e. culture medium, typically increases the background signal in the assay. For example, the CLuc assay solution alone shows 101–102 RLU; the CLuc assay solution in 10% FBS-containing medium results in 102–104 RLU, depending on the type of medium used.
The recommended integration time for the luminescence measurement is 2–10 seconds. It is important to keep the integration time constant, in order to obtain consistent results.